JPH06508156A - Protein preparation consisting of growth hormone - Google Patents

Protein preparation consisting of growth hormone

Info

Publication number
JPH06508156A
JPH06508156A JP5517378A JP51737893A JPH06508156A JP H06508156 A JPH06508156 A JP H06508156A JP 5517378 A JP5517378 A JP 5517378A JP 51737893 A JP51737893 A JP 51737893A JP H06508156 A JPH06508156 A JP H06508156A
Authority
JP
Japan
Prior art keywords
growth hormone
formulation
hgh
preparation
aqueous solution
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
JP5517378A
Other languages
Japanese (ja)
Other versions
JP3268557B2 (en
Inventor
カステンソン　ステファン
フロリン　ロバートソン　エバ
ホクビー　エルビー
トム　シルッカ
Original Assignee
ファーマシア　アクチボラーグ
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Family has litigation
First worldwide family litigation filed litigation Critical https://patents.darts-ip.com/?family=20385860&utm_source=***_patent&utm_medium=platform_link&utm_campaign=public_patent_search&patent=JPH06508156(A) "Global patent litigation dataset” by Darts-ip is licensed under a Creative Commons Attribution 4.0 International License.
Application filed by ファーマシア　アクチボラーグ filed Critical ファーマシア　アクチボラーグ
Publication of JPH06508156A publication Critical patent/JPH06508156A/en
Application granted granted Critical
Publication of JP3268557B2 publication Critical patent/JP3268557B2/en
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
    • A61K47/12—Carboxylic acids; Salts or anhydrides thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/27—Growth hormone [GH], i.e. somatotropin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Endocrinology (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Dermatology (AREA)
  • Zoology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Immunology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Medicinal Preparation (AREA)
  • Peptides Or Proteins (AREA)

Abstract

PCT No. PCT/SE93/00281 Sec. 371 Date Dec. 20, 1993 Sec. 102(e) Date Dec. 20, 1993 PCT Filed Apr. 1, 1993 PCT Pub. No. WO93/19776 PCT Pub. Date Oct. 14, 1993The invention relates to injectable formulations of growth hormone comprising citrate as buffer substance. The formulation could be an aqueous solution of growth hormone and citrate as buffer substance in a concentration of 2-50 mM. The formulation of the growth hormone could comprise growth factor, amino acids such as e.g. glycine and alanine and/or mannitol or other sugar alcohols and/or glycerol and/or other carbohydrates and optionally a preservative such as benzyl alcohol. The invention also relates to a process for preparation of the formulation by mixing growth hormone with citrate as buffer substance or by adding the constituents of the final formulation on the last gel purification step.

Description

【発明の詳細な説明】 発明の名称 成長ホルモンから成る蛋白調合物 技術分野 本発明は成長ホルモンもしくはその機能的類似体の調合物に関するものであり、 特に緩衝材料として濃度が約２乃至５０ｍＭのシトラード、とりわけ濃度が約２ 乃至２０ｒｎＭのクエン酸ナトリウムから成る人間の成長ホルモン（ｈＧＨ）溶 液に関するものである。この緩衝材料は安定性の向上に用いられる。[Detailed description of the invention] name of invention Protein preparation consisting of growth hormone Technical field The present invention relates to preparations of growth hormone or functional analogues thereof; In particular, as a buffer material citrad at a concentration of about 2 to 50 mM, especially at a concentration of about 2 A human growth hormone (hGH) solution consisting of 20rnM sodium citrate. It is related to liquid. This buffer material is used to improve stability.

背景技術 成長ホルモンはヒトと動物の双方のもの、たとえばヒト成長ホルモン（ｈＧＨ） 、ウシ成長ホルモン（ｂＨＧ）と、豚成長ホルモン（ｐＧＨ）である。Background technology Growth hormones are found in both humans and animals, such as human growth hormone (hGH) , bovine growth hormone (bHG), and porcine growth hormone (pGH).

））　Ｇ　Ｈは１９１のアミノ酸の単鎖から成る蛋白である。)) GH is a protein consisting of a single chain of 191 amino acids.

分子は２つのジスルフィド結合で架橋され、単量体フオームは２２ｋＤａの分子 ・量を有する。しかし、下垂体ヒト成長ホルモンは均質でない。たとえば、同一 遺伝子からつくられる比較的小さい２０　ｋ　Ｄ　ａ　ｈ　Ｇ　）ｌ変異体も周 知である。妊娠中の胎盤により示される「基礎ｈＧＨＪ変異体（ｈＧＨ−Ｖ）は もう１つの類似体であり、別の遺伝子の生成物である。２２ｋＤａ　ｈＧＨのよ うに、それは１９１のアミノ酸から成るが、それらの分子１３の全体を通して各 種の位置において異なっている。たとえば、１９７５年発行のビューレイ・チー ・ニー代著による「前進酵素学ｊ、第４２巻、第７３頁乃至第１６６頁及び１９ ８８年発行のフランケン・エフ代著による［内分泌及び新陳代謝臨床ジャーナル ］、第６６巻、第１１７１頁乃至第１１８０頁参照のこと。The molecule is cross-linked with two disulfide bonds, and the monomer form is a 22 kDa molecule. ・Have a quantity. However, pituitary human growth hormone is not homogeneous. For example, the same Relatively small 20kDahG)l mutants created from genes are also common. It is knowledge. The “basal hGHJ variant (hGH-V)” exhibited by the placenta during pregnancy is Another analogue, the product of another gene. 22kDa hGH It consists of 191 amino acids, but each throughout the molecule's 13 They differ in the location of the species. For example, Beaulay Chi, published in 1975. ・Advanced Enzymology J, Vol. 42, pp. 73 to 166 and 19 by Nie Dai [Clinical Journal of Endocrinology and Metabolism] by Franken F., published in 1988. ], Vol. 66, pp. 1171-1180.

組換えｈＧＨ（２２ｋＤａ）はここ数年の間に市販で入手できてきた。それはヒ トの組織から調製された生成物がクロイツエフエルト・ヤコブ病のような伝染性 病原体を含む虞があるので下垂体抽出生成物よりもむしろ好まれている。治療上 有用な２種類の組換えｈＧＨ調合剤が市場に出ている。権威あるものとしてはた とえば、カビ　ファーマシア　アクチボラーグ社のゲノトロピン（商標）と、Ｎ −末端で付加メチオニン残留物のある類似体、たとえばソマトノーム（商標）で ある。Recombinant hGH (22 kDa) has been commercially available for several years. That's Hi Products prepared from human tissue may cause infectious diseases such as Creutzefuelt-Jakob disease. It is preferred over pituitary extract products due to their potential to contain pathogens. therapeutically There are two useful recombinant hGH preparations on the market. as an authoritative For example, Genotropin (trademark) from Kabi Pharmacia Actiborag and N - Analogues with additional methionine residues at the end, e.g. in Somatonome™ be.

ｈＧＨは下垂体機能不全小人症またはターナ−症候群の患者の線形成長を刺激す るために用いられるが、他の適応症にも示唆されている。hGH stimulates linear growth in patients with hypopituitarism dwarfism or Turner syndrome. Although it is used for treatment, other indications have also been suggested.

水性化合物中の蛋白の安定性は一般に製薬業界において問題がある。それは各種 の乾燥法たとえば凍結乾燥による蛋白の乾燥により解決されてきた。それ以来、 蛋白は乾燥形式で販売されそして貯蔵されてきた。患者はその乾燥蛋白を止むを 得ず使用前に溶剤で元に戻す必要があり、これは患者にとってはもちろん不利か つ不便である。The stability of proteins in aqueous compounds is generally a problem in the pharmaceutical industry. It is various This problem has been solved by drying the protein by, for example, freeze-drying. since then, Protein has been sold and stored in dry form. The patient should stop taking the dry protein. Unfortunately, it must be reconstituted with a solvent before use, which is of course disadvantageous for the patient. It's inconvenient.

新しい投与装置、たとえばカビ　ファーマシア　アクチボラーグ社の米国特許第 ４，９６８，２９９号明細書に記載されたカビペン（商標）により、患者はどち らかと言えば取扱いの簡単な装置を与えた。この装置は２室のアンプルのゲノミ ックス（商標）から成り、その片方の室に凍結乾燥粉末としてのｈＧＨを入れ、 他方の室には元に戻す希釈液を入れである。患者は使用前にその製品を元に戻す 。凍結乾燥製品は最長２４ケ月の間貯蔵できる。元に戻した製品は２乃至８℃の 温度で貯蔵すると３週間は安定している。New dosing devices, such as Kabi Pharmacia Actiborag's U.S. Patent No. No. 4,968,299, the Kavipen (trademark) allows patients to To be honest, we have provided a device that is easy to handle. This device consists of a two-chamber ampoule genome. (trademark), one chamber of which contains hGH as a freeze-dried powder, The other chamber contains the diluent to be reconstituted. The patient should return the product before use. . Freeze-dried products can be stored for up to 24 months. The reconstituted product should be kept at a temperature of 2 to 8 degrees Celsius. Stable for 3 weeks when stored at room temperature.

凍結乾燥法は高価につく時間浪費の作業工程であり、蛋白の市販製品を調合する 際、この工程を避けることができる場合、それは非常に有利となる。Freeze-drying is an expensive and time-consuming process for formulating commercial products of proteins. It would be very advantageous if this step could be avoided.

成長ホルモン、たとえばｈ　Ｇ　）（の注射を毎日必要とする患者で、特にその 患者が子供である時、製品の取扱い、投薬ならびに注射が容易なことが重要であ る。凍結乾燥ｈ　Ｇ　Ｈの水戻しには慎重さと注意深さが必要で、なるべくなら 避けた方がよいが、今日唯一の利用できる方法である。Patients who require daily injections of growth hormone (e.g. hG), especially When the patient is a child, it is important that the product is easy to handle, administer, and inject. Ru. Rehydration of freeze-dried hGH requires caution and caution, and if possible, It is best avoided, but it is the only method available today.

蛋白を溶液として生産し、患者に販売でき、その患者が水に戻すことなく薬剤を 直接注射できる場合、成長ホルモン、特にｈ　Ｇ　Ｈの使用は容易になる。Proteins can be produced as a solution and sold to patients, who can administer the drug without having to put it back into the water. The use of growth hormone, particularly hGH, is facilitated if it can be directly injected.

この問題の色々な解決法が開示されてきたが、これまで製品として市場に出たも のはない。Various solutions to this problem have been disclosed, but so far none have reached the market as products. There is no.

ジェネンテック氏の国際特許出願公開第Ｗ○８９１０９６１４号公報では、グリ シンと、マンニトールと、緩衝剤とから成るｈＧＩＩの安定配合物を開示し、ま た好ましい実施例では、非イオン界面活性剤たとえばポリソルベート８０を添加 する。燐酸ナトリウムが緩衝物質として示唆されている。配合物は凍結乾燥調合 剤にした時、また水で戻した際も向上した安定性がある。In Genentech's International Patent Application Publication No. W○89109614, discloses a stable formulation of hGII consisting of mannitol, mannitol, and a buffer; In a preferred embodiment, a nonionic surfactant such as polysorbate 80 is added. do. Sodium phosphate has been suggested as a buffering substance. The formulation is a freeze-dried formulation. It has improved stability when made into a preparation and when reconstituted with water.

溶液にした成長ホルモンの投与の別の可能性はインターナショナル　ミネラルズ 　アンド　ケミカル社のヨーロッパ特許第２１１，６０１号明細書によればポリ オキシエチレン−ポリオキシプロピレンを含むグロック共重合体を添加すること である。この溶液は投与に際し、動物に持続性解放感を与える。Another possibility for administering growth hormone in solution is International Minerals. According to European Patent No. 211,601 by And Chemical Co., Ltd. Adding Glock copolymer containing oxyethylene-polyoxypropylene It is. Upon administration, this solution provides a sustained sensation of relief to the animal.

安定した、注射のできる成長ホルモン溶液、たとえば溶液にしたｈＧＨについて は市場での需要がある。最終製剤溶液が単に最低量に限った添加剤たとえばテン シトを含む場合も有利である。For stable, injectable growth hormone solutions, such as hGH in solution is in demand in the market. If the final formulation solution contains only minimal amounts of excipients, e.g. It is also advantageous if it contains carbon.

ここで上述した諸問題を解決する新しい配合物を見出した。We have now found a new formulation that solves the problems mentioned above.

発明の開示 ＧＨの安定性 ｈ　Ｇ　Ｉ（の安定性は蛋白の化学的ならびに物理的性質に左右される。異なる 分解径路、たとえば脱アミド化、酸化および凝集が周知である。Disclosure of invention Stability of GH h　G　I(stability) depends on the chemical and physical properties of the protein. Degradation pathways such as deamidation, oxidation and aggregation are well known.

脱アミド化と酸化は蛋白の一次構造の変化から成る普通の化学反応である。脱ア ミド化は特に水溶液で生じるが、水溶液の低温と低ｐ　ｔｉは脱アミド化反応を 抑制する。Deamidation and oxidation are common chemical reactions that involve changes in the primary structure of proteins. Departing from Africa Mididation occurs particularly in aqueous solutions, but the low temperatures and low pti of aqueous solutions inhibit the deamidation reaction. suppress.

凝集の異なる形態は蛋白の物理的不安定性に起因する。The different forms of aggregation are due to the physical instability of the protein.

凝集は可溶性にも不溶性にもなり得て、両形態の双方の結合は共有結合にも非共 有結合にもなり得る。凝集は乳光溶液をつくるが、化学的にのみ示し得る非可視 凝集と成る。蛋白配合物における共有凝集の防止は前記の方法が不可逆で、その うえ免疫原性となり得る不活性種が結果として生じることもあり得るので重要で ある。−次構造の変化は蛋白の自己会合である凝集の原因となり得る配座変化も 生じさせることがある。一定の条件で起こる非共有凝集は沈澱と活性の損失に導 くことがある。Aggregates can be soluble or insoluble, and both forms of binding can be covalent or non-covalent. It can also be a bond. Aggregation creates an opalescent solution, but it is invisible and can only be shown chemically. It becomes agglomeration. Prevention of covalent aggregation in protein formulations is achieved by the methods described above, which are irreversible and This is important because it can also result in inert species that can be immunogenic. be. -Changes in the next structure also include conformational changes that can cause aggregation, which is protein self-association. It may cause Non-covalent aggregation that occurs under certain conditions leads to precipitation and loss of activity. There are times when

多数の反応が異なるｐ　Ｈ条件下で起こり得るので、あらゆる変態反応を排除し ながら蛋白の高溶解度と固有の配座を維持させる特定のｐＨでの蛋白の配合はほ とんど不可能である。Since a large number of reactions can occur under different pH conditions, any transformation reactions should be excluded. However, formulation of the protein at a specific pH that maintains high solubility and unique conformation of the protein is very difficult. It's almost impossible.

今まで、僅かにアルカリ性のｐＨは一般に可視粒子の予防と透明薬品の達成のた め製造業者により利用されてきた。はとんどの市販製品において、ｐＩ−１は脱 アミド化の危険性が高いにもかかわらず７以上もある。Until now, a slightly alkaline pH was generally used to prevent visible particles and to achieve transparent drugs. has been used by many manufacturers. In most commercially available products, pI-1 is Despite the high risk of amidation, there are 7 or more.

カビ　ファーマシア　アクチボラーグ社の製品であるゲノトロビン（商標）を水 で戻す時、１６ＩＵ／ｍｌのｈＧＩｒ濃度て６．７というｐ　Ｈを達成できる。Genothrobin (trademark), a product of Kabi Pharmacia Actiborag, is added to water. When reconstituted, a pH of 6.7 can be achieved with an hGIr concentration of 16 IU/ml.

このｐ　）（は全く透明な溶液（ｐＨ８）をつくるｐＨと、いくぶん乳光が強い が比較的低い脱アミド化速度を示すｐＨ６との間の中間のものである。This p is intermediate between pH 6 and 6, which shows a relatively low deamidation rate.

この複雑さのため、蛋白製剤を配合することはできず、又全分解径路の排除も不 可能である。凍結乾燥蛋白製品は同様の水溶液よりもずっと安定している。しか し、凍結乾燥製品は加工後は十分安定しているが、蛋白はそれでも貯蔵中に緩慢 に分解する。Because of this complexity, it is not possible to formulate protein preparations and eliminate all degradation pathways. It is possible. Lyophilized protein products are much more stable than similar aqueous solutions. deer However, although freeze-dried products are stable enough after processing, proteins still stagnate during storage. Decompose into.

シトラードを緩衝物質として選んで入れた成長ホルモンを含む溶液が燐を緩衝剤 に入れたものよりもずっと安定していることがわかって全く意外であった。A solution containing growth hormone containing citrad as a buffering substance uses phosphorus as a buffering agent. I was completely surprised to find that it was much more stable than what I had put in it.

本発明は、緩衝物質としてのシトラードから成る成長ホルモンもしくはその機能 性類似体の注射できる調合物に関するものである。該調合物は成長ホルモンもし くはその類似体の水溶液であり、その中に緩衝物質としてシトラードを２乃至５ ０ｍＭの濃度で、また好ましくは緩衝物質としてクエン酸ナトリウムを約５．０ 乃至７．５のｐＨ１２乃至４０ｍＭの濃度で含有している。The present invention provides a growth hormone comprising citrad as a buffer substance or its function. The invention relates to injectable formulations of sex analogues. The preparation may contain growth hormone. is an aqueous solution of its analogue, in which citrad is added as a buffer substance. Sodium citrate at a concentration of 0mM and preferably as a buffer substance is about 5.0% It is contained at a concentration of 12 to 40mM at a pH of 12 to 7.5.

なるべくなら調合物はｈ　Ｇ　Ｈもしくはその機能性類似体と緩衝物質としての シトラードを２乃至２０ｍＭたとえば５ｍＭ乃至１０ｍＭの濃度の水溶液にした もの、また好ましくはクエン酸ナトリウムを緩衝物質として約６゜０乃至７．０ のｐ　Ｈで含有する。Preferably the formulation contains hGH or a functional analogue thereof and as a buffer substance. Citrade was made into an aqueous solution at a concentration of 2 to 20mM, for example 5mM to 10mM. from about 6°0 to 7.0%, preferably using sodium citrate as a buffering substance. Contains at a pH of

成長ホルモンもしくはその機能性類似体の調合物はアミノ酸、たとえばグリセリ ンとアラニンまたは（および）マンニトールあるいは他の糖アルコールまたは（ および）グリセロールまたは（および）他の炭水化物と、任意的な保存料たとえ ばベンジルアルコールとから構成することができる。その溶液はアイソトニック にする必要がある。成長ホルモンは好ましくは再結合ｈ　Ｇ　Ｈである。Preparations of growth hormone or its functional analogues contain amino acids such as glycerin. and alanine or (and) mannitol or other sugar alcohols or ( and) glycerol or (and) other carbohydrates and optional preservatives. and benzyl alcohol. the solution is isotonic It is necessary to The growth hormone is preferably recombined hGH.

本発明による調合物は少なくとも１２ケ月間安定している。ここで安定というこ とは８５％以−Ｌの単量体（■ＥＦ＞と、２％以下の５ＤＳ−ＰＡＧＥによる断 片の量を意味する。Formulations according to the invention are stable for at least 12 months. What is stable here? means 85% or more of the monomer (■EF>) and 2% or less of 5DS-PAGE. means the amount of pieces.

本発明の調合物はさらに成長ホルモンと成長因子の混合物から構成することもで きる。成長因子はインシュリンの様な成長因子（ＩＧＦ−１またはＩＧＦ−２） と、表皮性成長因子（Ｅ　ＧＦ　）で、天然材料もしくは再結合技術による生成 物のいずれかを意味する。The formulation of the invention may further consist of a mixture of growth hormone and growth factors. Wear. Growth factors are insulin-like growth factors (IGF-1 or IGF-2) and epidermal growth factor (EGF), which can be produced from natural materials or by recombination technology. It means any of the things.

本発明はさらに成長ホルモンもしくはその機能性類似体を緩衝物質としてのシト ラードに混合させることにより、あるいは最終配合物の成分を最終ゲル精製工程 で添加することにより調合物を調剤する方法に関する。また、成長ホルモンもし くはその機能性類似体を必要とする患者に調合物を投与して処置する方法に関す るものである。The present invention further provides the use of growth hormone or a functional analogue thereof as a buffer substance. By mixing with lard or the components of the final formulation into the final gel purification step A method of preparing a formulation by adding . Also, growth hormone or a functional analogue thereof, for administering the formulation to a patient in need thereof. It is something that

成長ホルモン（ＧＨ）は天然に産出するヒトと動物のＧＩ−■と再結合Ｇ）−（ （ｒＧＨ）　、たとえばｒｈＧＨ，ｒｂＧＨとｒ　ｐ　Ｇ　）（の双方を言う。Growth hormone (GH) recombines with naturally occurring human and animal GI-■ (rGH), for example rhGH, rbGH and rpG).

機能性類似体は動物とヒトの成長ホルモンと同一の治療効果をもつ調合物を言う 。Functional analogues refer to preparations that have the same therapeutic effect as growth hormone in animals and humans. .

緩衝剤に含まれる成長ホルモンは初期単離生成物もしくは凍結乾燥後、戻した製 品であっても差し支えない。The growth hormone contained in the buffer may be the initial isolated product or the reconstituted product after lyophilization. There is no problem even if it is a product.

成長ホルモンの濃度は使用ずみ緩衝剤の安定性と、所定の投薬の所望治療量にの み左右される。なるべくならｋ］Ｇ　Ｈの濃度は１乃至８０ＩＵ／ｍｌで、さら に好ましくは２乃至４０１Ｕ／ｍｌである。The concentration of growth hormone depends on the stability of the buffer used and the desired therapeutic dose for a given medication. It depends on you. Preferably, the concentration of [k]GH is between 1 and 80 IU/ml, and It is preferably 2 to 401 U/ml.

溶液を振動させることにより観測される沈澱の形態にある凝縮は、空気の液界面 における変成の結果である。Condensation in the form of precipitation, observed by vibrating a solution, occurs at the air-liquid interface. is the result of metamorphosis in

これは充填空隙を残さず充填することで発生が防止でき、従って凝集の条件を減 少させることができる。This can be prevented by filling without leaving any voids, thus reducing the conditions for agglomeration. It can be made smaller.

発明を実施するための最良の形態 実施例１ 配合研究の材料は普通のゲノトロピン（商標）の方法から得られた。ヒトの成長 ホルモンはバクテリアである大腸菌（Ｅｃｏｌｉ）Ｋ１２において形質発現プラ スミドｐＡＰＳＴｌ　１　ｈＧＨ−３をテンプレートとして用いて合成する。前 記ホルモンは合成中に細胞から原形質膜間隙に分泌される。ヒト成長ホルモンは その後、引続いて、外側バクテリア膜の***の後に、前記Ｅｃｏ　ｌ　ｉ原形質 膜間隙から分離される。再結合ｈＧＨを含む抽出物ＤＥＡＥ−セファローズＦＦ で分別した。２つの硫酸アンモニウム沈澱工程がつづき、その後、ＤＥＡＥ−セ ファローズＦＦで２つの分別工程を行った。先の精製工程に用いられた塩の除去 の目的に役立つゲル濾過と、最終配合成分の添加により配合が行われた。最終分 離管セファデックスＧ−２５（直径１．３ｃｍ、ヘッド高さ４５ｃｍ）を配合緩 衝剤で平衡させた。平衡とクロマトグラフィーを＋７℃の温度で行った。所望の 蛋白濃度を配合緩衝剤で希釈することにより達成できた。７つの溶液の安定性を 調査した。表１参照のこと。BEST MODE FOR CARRYING OUT THE INVENTION Example 1 Materials for formulation studies were obtained from conventional Genotropin™ methods. human growth Hormones play a role in gene expression in the bacterium Escherichia coli (Ecoli) K12. Sumido pAPSTl 1 is synthesized using hGH-3 as a template. Before During synthesis, these hormones are secreted from cells into the plasma membrane space. human growth hormone is Subsequently, after division of the outer bacterial membrane, the Ecoli plasma Separated from the intermembrane space. Extract DEAE-Sepharose FF containing recombined hGH It was separated by Two ammonium sulfate precipitation steps follow, followed by DEAE-separation. Two fractionation steps were performed with Fallows FF. Removal of salts used in previous purification steps The formulation was carried out by gel filtration which served the purpose of and addition of the final formulation ingredients. final minute Contains sephadex G-25 (diameter 1.3cm, head height 45cm). Equilibrated with buffer. Equilibration and chromatography were carried out at a temperature of +7°C. desired Protein concentration could be achieved by diluting with formulation buffer. Stability of 7 solutions investigated. See Table 1.

表１ 実施例　ＡＢＣＤ ｈＧＨＩＵ／ｍｌ　２０　２０　１０　１０クエン酸ナトリウムｍＭ　５　−　 − 燐酸ナトリウムｒｎＭ　５　１０　１０グリシンｍＭ　１２　１２　１２．　１ ２マンニド−／ＩｚｍＭ　２５０　２５０　２５０　２５０ｐ　ト１　６．２　 ６．３　６．２　７．４容積　１１１１ 出発値： ｐＨ６，２６，３６，２７，４ ＩＥＦ（％単量体）　９８　９８　９９　１００ＳＤＳ−ＤＡＧＥ 凝縮％　００−− 単量体％　９９．８　９９．６　− 断片％　０．２　０．４　−　− 可視検査　透明　透明　透明　透明 ５℃の温度で貯蔵６ケ月後の結果 ｐＨ６，３６，４６，４７，５ ＩＥＦ　（％単量体）　８８　８４　８８　７３ＳＤＳ−ＤＡＧＥ 凝縮％　ＯＯＯＯ，２ 単量体％　９９．７　９５．１　９７．５　９８．４断片％　０．３　４．９　 ２．５　１．３可視検査　透明 ５℃の温度で貯蔵１５ケ月後の結果 ｐＨ６，３６，４ ＩＥＦ（％単量体）　８６　７５ ＳＤＳ−ＤＡＧＥ 凝縮％　００．１ 単量体％　９８．９　９２．７ 断片％　１．０　７．３ 可視検査　透明　透明 ５℃の温度で貯蔵２４ケ月後の結果 ｐＨ６，４６，４ ＩＥＦ　（％単量体）　８８　７１ ＳＤＳ−ＤＡＧＥ 凝縮％　００ 単量体％　９８．３　８９．１ 断片％　１，７　１０．９ 可視検査　透明　透明 表２ 実施例　ＥＦＧＨ ｈＧＨＩＵ／ｍｌ　４　４　１０　１０ク工ン酸ナトリウムｒｒｔＭ　１０　〜 　−　５燐酸ナトリウムｍＭ　−１０１０− グリシンｍＭ　１２　１２　１２　１２マンニト一ルｍＭ　２５０　２５０　２ ５０　２５０ｐＨ６，２６，１６，３６，１ 容積　３　３　２０．３５ 出発値： ｐＩｉ　６．２　６．１　６．３　６．ＩＩＥＦ（％単量体）　９９　９９　９ ７　１００可視検査　透明　透明　透明　透明 ５℃の温度で貯蔵６ケ月後の結果 ｐＨ６，３６，２６，３６，６ ＩＥＦ（％単量体）　８９　８６　８５　９１ＳＤＳ−ＤＡＧＥ 凝縮％　０　０　０．１　０ 単量体％　９９．８　９７，１　９５，２　９８．１断片％　０．２　２．９　 ４．７　１．９可視検査　透明　透明　透明　透明 ５℃の温度で貯蔵１２ケ月後の結果 ｐｉ（６，３６，５ Ｉ　ＩＥ　Ｆ（％単量体）　７１　９０ＳＤＳ−ＤＡＧＥ 凝縮％　０．４　０ 単量体％　９３　９８．１ 断片％　６．６　１．８ 可視検査　透明 方法 濃度Ｊ１測定による等電集束法（ＩＥＦ）Ｉ　Ｅ　Ｆは脱アミド化の程度が測定 できる方法である。Table 1 Example ABCD hGHIU/ml 20 20 10 10 Sodium citrate mM 5 - − Sodium phosphate rnM 5 10 10 Glycine mM 12 12 12. 1 2 Mannide/IzmM 250 250 250 250p 1 6.2 6.3 6.2 7.4 Volume 1111 Starting value: pH6,26,36,27,4 IEF (% monomer) 98 98 99 100 SDS-DAGE Condensation% 00-- Monomer% 99.8 99.6 - Fragment% 0.2 0.4 - - Visual inspection Transparent Transparent Transparent Transparent Results after 6 months storage at 5℃ temperature pH6,36,46,47,5 IEF (% monomer) 88 84 88 73 SDS-DAGE Condensation% OOOO, 2 Monomer% 99.7 95.1 97.5 98.4 Fragment% 0.3 4.9 2.5 1.3 Visual inspection Transparent Results after 15 months of storage at a temperature of 5℃ pH6,36,4 IEF (% monomer) 86 75 SDS-DAGE Condensation% 00.1 Monomer% 98.9 92.7 Fragment% 1.0 7.3 Visual inspection Transparent Transparent Results after 24 months storage at 5℃ temperature pH6,46,4 IEF (% monomer) 88 71 SDS-DAGE Condensation% 00 Monomer% 98.3 89.1 Fragment% 1,7 10.9 Visual inspection Transparent Transparent Table 2 Example EFGH hGHIU/ml 4 4 10 10 Sodium citrate rrtM 10 ~ - Sodium 5-phosphate mM -1010- Glycine mM 12 12 12 12 Mannitol mM 250 250 2 50　250pH6,26,16,36,1 Volume 3 3 20.35 Starting value: pIi 6.2 6.1 6.3 6. IIEF (% monomer) 99 99 9 7 100 visible inspection transparent transparent transparent transparent Results after 6 months storage at 5℃ temperature pH6,36,26,36,6 IEF (% monomer) 89 86 85 91 SDS-DAGE Condensation% 0 0 0.1 0 Monomer% 99.8 97.1 95.2 98.1 Fragment% 0.2 2.9 4.7 1.9 Visual inspection Transparent Transparent Transparent Transparent Results after 12 months of storage at a temperature of 5℃ pi(6,36,5 IIE F (% monomer) 71 90SDS-DAGE Condensation% 0.4 0 Monomer% 93 98.1 Fragment% 6.6 1.8 Visual inspection transparent Method Isoelectric focusing method (IEF) by concentration J1 measurement IEF measures the degree of deamidation This is a possible method.

２つの電極の間で設定され且つキャリヤーの両性電解質により安定するｌ）　Ｈ 勾配でｈ　Ｇ　Ｈ成分の分離を行う。蛋白は、蛋白が勾配のその等電点でそれ自 体整合するまで移行し、その等電点においては蛋白に正味総合電荷を帯びず、従 って移行が終るに従って凝縮する。従って、分離は電荷により達成される。帯電 ｈＧＨの形の相対的分布をクーマツシーブルー染色ポリペプチドの濃度計走査に より計算する６単量体の割合が高ければ高いほど脱アミド化は少なくなる。l) set between two electrodes and stabilized by the carrier ampholyte Separate the hGH component using a gradient. The protein will self-isolate at its isoelectric point of the gradient. At that isoelectric point, the protein carries no net overall charge and is It condenses as the transition ends. Separation is therefore achieved by charge. electrification Relative distribution of hGH forms in densitometric scans of Coomassie blue-stained polypeptides The higher the calculated proportion of 6 monomers, the less deamidation.

ポリペプチド大きさの分布（ＳＤＳ−ＰＡＧＥ）ソマトトロピンｈＧＨの調剤に おいて蛋白をドデシル硫酸ナトリウム（ＳＤＳ）により変成させて５ＤＳ−蛋白 の陰電荷分子鎖体を産出する。その後、分子の大きさによる分離をＳＤＳの共存 においてポリアクリルアミドゲル（ＰＡＧＥ）中における電気泳動により達成し た。Polypeptide size distribution (SDS-PAGE) for the preparation of somatotropin hGH The protein was denatured with sodium dodecyl sulfate (SDS) to obtain 5DS-protein. produces negatively charged molecular chains. After that, separation by molecular size was performed using SDS. achieved by electrophoresis in polyacrylamide gels (PAGE) in Ta.

ｈ　Ｇ　Ｈの相対的ポリペプチドの大きさによる分布を銀染ポリペプチドバンド の濃度計走査により計量した。h　G　H distribution according to relative polypeptide size as silver-stained polypeptide band densitometer scanning.

可視検査 溶液の外観をｐｈ、Ｅｕｒ第２版により目視検査を行った。visual inspection The appearance of the solution was visually inspected using pH and Eur 2nd edition.

ｐＨ ｐＨをガラスとカロメルの電極を用いて測定した。pH pH was measured using glass and calomel electrodes.

実施例Ａ、ＥとＨは本発明によるものである。Examples A, E and H are according to the invention.

表１と２から、断片の百分率は燐酸ナトリウムで緩衝された溶液中ではクエン酸 ナトリウムによるよりもずっと高いことが明らかである。Ｂ（燐酸ナトリウム） には５℃の温度で１５ケ月で７．３％の断片と９２．７％の単量体が含まれ、Ａ （クエン酸ナトリウム）にはそれぞれ１．０％と９８．９％が含まれている。From Tables 1 and 2, the percentage of fragments in the sodium phosphate buffered solution is It is clear that it is much higher than that due to sodium. B (sodium phosphate) contains 7.3% fragments and 92.7% monomer after 15 months at a temperature of 5°C, A (Sodium citrate) contains 1.0% and 98.9%, respectively.

Ｆ（燐酸ナトリウム）の断片の相対量は２．９％、単量体の百分率が５℃の温度 で６ケ刀後で９７．１％である。Ｅ（クエン酸すｌ・リウム）にはそれぞれ０． 　２％と９９．８％である。Ｂ、Ｃ，Ｄ、ＦとＧには燐酸ナトリウムが含まれ、 すべてに高い比率の断片が貯蔵後にも含まれている。Ｄは高いｐ　Ｈを有するが 、断片の量は１０ｍＭのクエン酸ナトリウム緩衝剤を用いる溶液Ｅのものよりも 高い。脱アミド化のグレードはＤにおいて許容できないほど高い。The relative amount of fragments of F (sodium phosphate) is 2.9%, the percentage of monomer is at a temperature of 5 °C After 6 swords, it is 97.1%. E (sl/lium citrate) has 0. 2% and 99.8%. B, C, D, F and G contain sodium phosphate, All contain a high proportion of fragments even after storage. Although D has a high pH , the amount of fragments is greater than that of solution E using 10 mM sodium citrate buffer. expensive. The grade of deamidation is unacceptably high in D.

実施例２ この実施例はベンジルアルコールを含む本発明による組成物とそれを含まないも のを比較するため行った。表３参照のこと。Example 2 This example shows compositions according to the invention containing benzyl alcohol and those without it. I went there to compare. See Table 3.

調合物を実施例１で説明したものと同一の方法で調剤したが、調合緩衝液にはベ ンジルアルコールがふくまれていた。The formulation was formulated in the same manner as described in Example 1, except that the formulation buffer Contains alcohol.

表３ 実施例　ＩＫＬＡ ｈＧＨＩＵ／ｍｌ　２０　２０　２０　２０ク工ン酸ナトリウムｍＭ　１０　１ ０　５　５グリシンｍＭ　１２　−　１２　１２ マンニト一ルｍＭ　１５０　１５０　１３０　２５０ベンジルアルコール％　１ 　１　１− ｐＨ６，３６，３６，３６，２ 容積　１　１　３．５　１ ３０℃の温度で貯蔵３週間後の結果 ｐＨ６，３６，３６，２６，２ ＩＥＦ　（％単量体）　６７　６８　６４　７２透明度　透明　透明　透明　透 明 ５℃の温度で貯蔵１ケ月後の結果 ｐＨ６，３６，３６，２６，２ ＩＥＦ　（％単量体）　９９　９９　−　９７透明度　透明　透明　透明　透明 ５℃の温度で貯蔵３ケ月後の結果 ｐＨ６，３６，４６，２６，３ ＩＥＦ　（％単量体）　９４　９４　９６　９４透明度　透明　透明　透明　透 明 ５ＤＳ−ＤＡＧＥ 凝縮％　ｏ　ｏ、ｉ 単量体％　９９．８　９９．４ 断片％　０．２　０．５ 実施例２はベンジルアルコールの添加がクエン酸ナトリウムを緩衝剤として用い た場合、安定性にどのような影響を及ぼすかを示すために行った。ベンジルアル コールは注射できる多投薬調剤としての使用に条件づきの保存料である。薬局方 によれば、適当な保存料を注射できる多投薬調剤に添加して製品の微生物上の安 全性の保証をめられている。Table 3 Example IKLA hGHIU/ml 20 20 20 20 Sodium citrate mM 10 1 0 5 5 glycine mM 12 - 12 12 Mannitol mM 150 150 130 250 Benzyl alcohol% 1 1 1- pH6,36,36,36,2 Volume 1 1 3.5 1 Results after 3 weeks of storage at a temperature of 30°C pH6,36,36,26,2 IEF (% monomer) 67 68 64 72 Transparency Transparent Transparent Transparent Transparent Akira Results after 1 month storage at 5℃ temperature pH6,36,36,26,2 IEF (% monomer) 99 99 - 97 Transparency Transparent Transparent Transparent Transparent Results after 3 months storage at 5℃ temperature pH6,36,46,26,3 IEF (% monomer) 94 94 96 94 Transparency Transparent Transparent Transparent Transparent Akira 5DS-DAGE Condensation%　o　o,i Monomer% 99.8 99.4 Fragment% 0.2 0.5 Example 2 shows that the addition of benzyl alcohol uses sodium citrate as a buffering agent. This was done to show how the stability would be affected if benzyl al Kohl is a preservative subject to use as an injectable multi-dose preparation. Pharmacopoeia According to A guarantee of integrity is required.

ベンジルアルコールは　強性に影響を有するので、ベンジルアルコールが添加さ れた際に、マンニトールの量が調節された。Benzyl alcohol has a strong effect, so benzyl alcohol is not added. The amount of mannitol was adjusted accordingly.

表３からベンジルアルコールの添加がクエン酸を緩衝剤として用いる時、安定性 に影響を及ぼさないことが明らかである。Table 3 shows that the addition of benzyl alcohol improves stability when citric acid is used as a buffer. It is clear that there is no effect on

実施例３ この実施例は、グリシンおよびマンニトールを含む本発明の組成物と、それを含 まないものを比較するために行った。表４参照のこと。Example 3 This example describes a composition of the invention containing glycine and mannitol and a composition containing the same. I went there to compare what I don't have. See Table 4.

調合物を実施例１で説明したものと同一の方法で調合した。The formulation was prepared in the same manner as described in Example 1.

表４ 実施例　ＭＮ ｈＧＨＩＵ／ｍｌ　２０　２０ ク工ン酸ナトリウムｍＭ　５　５ グリシンｍＭ　−１２ マンニト一ルｍＭ　−１５０ ｐＨ６，３６，３ 容積　１１ ３０°Ｃの温度で貯蔵１週間後の結果 ｐＪ−（６，３６，３ ＩＥＦ（％単量体）　９２　９１ ＳＤＳ−ＤＡＧＥ 凝縮％　ＯＯ 単量体％　９９．５　９９．６ 断片％　０．５　０．４ 可視検査　透明　透明 表４から、添加剤たとえばグリシンとマンニトールの添加がクエン酸を緩衝剤と して用いる時、安定性に影響を及ぼさないことがわかる。Table 4 Example MN hGHIU/ml 20 20 Sodium citrate mM 5 5 Glycine mM -12 Mannitol mM -150 pH6,36,3 Volume 11 Results after one week of storage at a temperature of 30°C pJ-(6,36,3 IEF (% monomer) 92 91 SDS-DAGE Condensation%　OO Monomer% 99.5 99.6 Fragment% 0.5 0.4 Visual inspection Transparent Transparent From Table 4, it can be seen that the addition of additives such as glycine and mannitol makes citric acid a buffer. It can be seen that there is no effect on stability when used as a compound.

フロントページの続き （７２）発明者　ホクピー　エルビー スウェーデン国、ニス−１２２４６エンスケド、シックリンスバーゲン　６ （７２）発明者　トム　シルツカ スウェーデン国、ニス−１４３００パルビー、エル　ジー　３６０　マイルスタ グバーゲン　３５９Continuation of front page (72) Inventor Hokupi L.B. Schicklinsbergen, Nis-12246 Ensked, Sweden 6 (72) Inventor Tom Siltzka Sweden, Nis-14300 Palby, L.G. 360 Milesta Gubergen 359

Claims (1)

【特許請求の範囲】 １）緩衝物質として２乃至５０ｍＭの量のクエン酸から成るヒトの成長ホルモン 又はその機能性類似体の注射可能な調合物。 ２）調合物が成長ホルモンもしくはそれの機能性類似体の水溶液であることを特 徴とする請求の範囲第１項に記載の調合物。 ３）調合物がｈＧＨの水溶液であることを特徴とする請求の範囲第２項に記載の 調合物。 ４）調合物がｈＧＨと濃度２乃至２０ｍＭの緩衝物質としてのシトラートの水溶 液であることを特徴とする請求の範囲第３項に記載の調合物。 ５）緩衝物質として、ｐＨが約６乃至７のクエン酸ナトリウムから成ることを特 徴とする請求の範囲第２項乃至第４項のいずれか１項に記載のｈＧＨの水溶液。 ６）調合物が５ｍＭの量のクエン酸ナトリウムから成ることを特徴とする請求の 範囲第５項に記載のｈＧＨの水溶液。 ７）調合物が１０ｍＭの量のクエン酸ナトリウムから成ることを特徴とする請求 の範囲第５項に記載のｈＧＨの水溶液。 ８）調合物が１つ以上の数個の成長因子から成ることを特徴とする請求の範囲第 １項もしくは第２項に記載の成長ホルモンまたはその機能性類似体の調合物。 ９）調合物がグリシンと、マンニトールと、グリセロールまたはそのいずれかか ら成ることを特徴とする請求の範囲第１項、第２項又は第８項のいずれか１項に 記載の成長ホルモンまたはその機能性類似体の調合物。 １０）調合物が保存料から成ることを特徴とする請求の範囲第１項乃至第９項の いずれか１項に記載の成長ホルモンあるいはその機能性類似体の調合物。 １１）ホルモンがｈＧＨの水溶液であり、また保存料がベンジルアルコールであ ることを特徴とする請求の範囲第１０項に記載の成長ホルモン。 １２）ｈＧＨが再結合ｈＧＨであることを特徴とする請求の範囲第３項に記載の 成長ホルモンの調合物。 １３）成長ホルモンもしくはその機能性類似体を緩衝物質としてのシトラートと 混合するか、あるいは最終配合の成分を最終ゲル精製工程で添加することを特徴 とする請求の範囲第１項乃至第１２項のいずれか１項に記載の調合物の調合方法 。 １４）成長ホルモンもしくはその機能性類似体を必要とする患者を請求の範囲第 １項乃至第１２項のいずれか１項による配合を投与して治療する方法。[Claims] 1) Human growth hormone consisting of citric acid in an amount of 2 to 50 mM as a buffer substance or an injectable formulation of a functional analog thereof. 2) Specifying that the preparation is an aqueous solution of growth hormone or a functional analogue thereof; A formulation according to claim 1, characterized in that: 3) according to claim 2, characterized in that the preparation is an aqueous solution of hGH. concoction. 4) The preparation consists of an aqueous solution of hGH and citrate as a buffer substance at a concentration of 2 to 20 mM. 4. A formulation according to claim 3, characterized in that it is a liquid. 5) The buffer substance consists of sodium citrate having a pH of about 6 to 7. The aqueous solution of hGH according to any one of claims 2 to 4. 6) Claims characterized in that the formulation consists of sodium citrate in an amount of 5mM. Aqueous solution of hGH according to scope 5. 7) Claim characterized in that the formulation consists of sodium citrate in an amount of 10 mM An aqueous solution of hGH according to item 5. 8) Claim 1, characterized in that the preparation consists of one or more several growth factors. A preparation of growth hormone or a functional analogue thereof according to paragraph 1 or paragraph 2. 9) Whether the formulation contains glycine, mannitol, and/or glycerol Claims 1, 2 or 8, characterized in that: A preparation of the growth hormone or functional analog thereof as described. 10) Claims 1 to 9, characterized in that the preparation consists of a preservative. A preparation of growth hormone or a functional analogue thereof according to any one of the preceding clauses. 11) The hormone is an aqueous solution of hGH and the preservative is benzyl alcohol. The growth hormone according to claim 10, characterized in that: 12) The hGH according to claim 3, wherein the hGH is recombined hGH. Growth hormone preparations. 13) Growth hormone or its functional analogue with citrate as a buffer substance Characterized by mixing or adding the components of the final formulation in the final gel purification step A method for preparing a formulation according to any one of claims 1 to 12. . 14) Patients in need of growth hormone or its functional analogues A method of treatment by administering a combination according to any one of paragraphs 1 to 12.
JP51737893A 1992-04-03 1993-04-01 Protein formulation consisting of growth hormone Expired - Lifetime JP3268557B2 (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
SE9201073-5 1992-04-03
SE9201073A SE9201073D0 (en) 1992-04-03 1992-04-03 PROTEIN FORMULATION
PCT/SE1993/000281 WO1993019776A1 (en) 1992-04-03 1993-04-01 Protein formulation comprising growth hormone

Publications (2)

Publication Number Publication Date
JPH06508156A true JPH06508156A (en) 1994-09-14
JP3268557B2 JP3268557B2 (en) 2002-03-25

Family

ID=20385860

Family Applications (1)

Application Number Title Priority Date Filing Date
JP51737893A Expired - Lifetime JP3268557B2 (en) 1992-04-03 1993-04-01 Protein formulation consisting of growth hormone

Country Status (15)

Country Link
US (1) US5567677A (en)
EP (1) EP0587858B2 (en)
JP (1) JP3268557B2 (en)
AT (1) ATE196092T1 (en)
AU (1) AU666007B2 (en)
CA (1) CA2102693C (en)
DE (1) DE69329367T3 (en)
DK (1) DK0587858T3 (en)
ES (1) ES2150940T5 (en)
GR (1) GR3034925T3 (en)
NO (1) NO310804B1 (en)
NZ (1) NZ251498A (en)
PT (1) PT587858E (en)
SE (1) SE9201073D0 (en)
WO (1) WO1993019776A1 (en)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6150331A (en) * 1998-02-04 2000-11-21 Jcr Pharmaceuticals Co., Ltd. Human growth hormone-containing aqueous pharmaceutical composition
JP2012051891A (en) * 1999-07-12 2012-03-15 Sandoz Ag Growth hormone formulation
JP2015514704A (en) * 2012-03-30 2015-05-21 ハンミ サイエンス カンパニー リミテッドＨａｎｍｉ Ｓｃｉｅｎｃｅｃｏ．，Ｌｔｄ． High concentration solution of long-acting human growth hormone conjugate

Families Citing this family (53)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5981485A (en) * 1997-07-14 1999-11-09 Genentech, Inc. Human growth hormone aqueous formulation
US5763394A (en) * 1988-04-15 1998-06-09 Genentech, Inc. Human growth hormone aqueous formulation
US5849700A (en) * 1991-12-20 1998-12-15 Novo Nordisk A/S Pharmaceutical formulation
US6022858A (en) * 1991-12-20 2000-02-08 Novo Nordisk A/S Pharmaceutical formulation of human-growth hormone pretreated with zinc salt
DK204791D0 (en) * 1991-12-20 1991-12-20 Novo Nordisk As UNKNOWN PHARMACEUTICAL PREPARATION
US5849704A (en) * 1991-12-20 1998-12-15 Novo Nordisk A/S Pharmaceutical formulation
FR2686899B1 (en) 1992-01-31 1995-09-01 Rhone Poulenc Rorer Sa NOVEL BIOLOGICALLY ACTIVE POLYPEPTIDES, THEIR PREPARATION AND PHARMACEUTICAL COMPOSITIONS CONTAINING THEM.
FR2719479B1 (en) * 1994-05-04 1996-07-26 Sanofi Elf Stable lyophilized formulation comprising a protein: assay kit.
IL115592A (en) * 1994-10-13 2000-07-26 Novo Nordisk As Stabilized pharmaceutical formulation comprising a growth hormone and leucine
US5631225A (en) * 1994-10-13 1997-05-20 Novo Nordisk A/S Pharmaceutical formulation
US5547696A (en) * 1994-10-13 1996-08-20 Novo Nordisk A/S Pharmaceutical formulation
US5654278A (en) * 1994-10-13 1997-08-05 Novo Nordisk A/S Composition and method comprising growth hormone and leucine
US5705482A (en) * 1995-01-13 1998-01-06 Novo Nordisk A/S Pharmaceutical formulation
US5552385A (en) * 1995-06-05 1996-09-03 Novo Nordisk A/S Pharmaceutical formulation
DE19539574A1 (en) 1995-10-25 1997-04-30 Boehringer Mannheim Gmbh Preparations and processes for stabilizing biological materials by means of drying processes without freezing
GB9526733D0 (en) * 1995-12-30 1996-02-28 Delta Biotechnology Ltd Fusion proteins
AUPN801296A0 (en) 1996-02-12 1996-03-07 Csl Limited Stabilised growth hormone formulation and method of preparation thereof
US20070179096A1 (en) 1996-04-24 2007-08-02 Novo Nordisk A/S Pharmaceutical Formulation
ZA9711733B (en) 1996-12-31 1998-07-01 Monsanto Co Method for solubilization and naturation of somatotropins
ZA9711731B (en) * 1996-12-31 1998-07-01 Monsanto Co Aqueous glycerol formulations of somatotropin
US6979442B1 (en) 1998-08-17 2005-12-27 Pfizer Inc. Stabilized protein compositions
TWI283182B (en) * 2000-08-07 2007-07-01 Nektar Therapeutics Inhalable spray dried 4-helix bundle protein powders having minimized aggregation
GB2371227A (en) * 2001-01-10 2002-07-24 Grandis Biotech Gmbh Crystallisation - resistant aqueous growth hormone formulations
WO2002092619A2 (en) * 2001-05-14 2002-11-21 The Gouvernment Of The United States Of America, Represented By The Secretary, Department Of Health And Human Services Modified growth hormone
EP1463752A4 (en) 2001-12-21 2005-07-13 Human Genome Sciences Inc Albumin fusion proteins
CA2475953A1 (en) * 2002-02-27 2003-09-04 E. Ann Tallant Angiotensin-(1-7) and angiotensin-(1-7) agonists for inhibition of cancer cell growth
CN1668332A (en) * 2002-07-09 2005-09-14 桑多斯股份公司 Liquid formulations with a high concentration of human growth hormone (HGH) comprising glycine
US20070065469A1 (en) * 2002-07-09 2007-03-22 Michael Betz Liquid formulations with high concentration of human growth hormone (high) comprising glycine
WO2004082707A2 (en) * 2003-03-18 2004-09-30 Ares Trading Sa Stabilisation of growth hormones in solution
WO2005063298A1 (en) 2003-12-23 2005-07-14 Pharmacia Corporation Stable growth hormone liquid formulation
UA89781C2 (en) * 2004-04-07 2010-03-10 Арес Трейдінг С.А. Liquid growth hormone formulation
EP1781700B1 (en) 2004-08-03 2014-03-19 TransTech Pharma, LLC Rage fusion proteins and methods of use
KR20070057818A (en) * 2004-08-03 2007-06-07 트랜스테크 파르마, 인크. Rage fusion proteins and methods of use
JP4951344B2 (en) * 2004-08-24 2012-06-13 第一三共株式会社 Bioactive peptide liquid formulation
ATE408624T1 (en) * 2004-11-09 2008-10-15 Ares Trading Sa METHOD FOR PURIFYING FSH
US20070015689A1 (en) * 2005-06-23 2007-01-18 Alza Corporation Complexation of metal ions with polypeptides
CN101282990B (en) * 2005-08-26 2013-04-03 阿雷斯贸易股份有限公司 Method for preparing glycosylated interferon-beta
EP1948221B1 (en) * 2005-11-02 2015-04-08 Syneron Medical Ltd. Human growth hormone patch formulations
JP5199112B2 (en) * 2005-12-09 2013-05-15 アレス トレーディング ソシエテ アノニム Method for purifying FSH or FSH variants
ES2431632T3 (en) * 2005-12-23 2013-11-27 Gcoder Systems Ab Positioning pattern
US20090004190A1 (en) * 2006-02-09 2009-01-01 Mjalli Adnan M M Rage Fusion Proteins And Methods Of Use
EP2380983A3 (en) * 2006-05-05 2012-12-05 TransTech Pharma Inc. RAGE fusion proteins, formulations, and methods of use thereof
WO2008100470A2 (en) * 2007-02-15 2008-08-21 Transtech Pharma, Inc. Rage - immunoglobulin fusion proteins
CA2687141C (en) 2007-05-22 2014-04-01 Amgen Inc. Compositions and methods for producing bioactive fusion proteins
WO2009053360A1 (en) * 2007-10-22 2009-04-30 Merck Serono S.A. Method for purifying an fc-containing protein
AU2008314687A1 (en) * 2007-10-22 2009-04-30 Merck Serono S.A. Method for purifying Fc-fusion proteins
WO2010122460A1 (en) 2009-04-20 2010-10-28 Pfizer Inc. Control of protein glycosylation and compositions and methods relating thereto
EP2427215A1 (en) * 2009-05-07 2012-03-14 Novo Nordisk Health Care AG Stable pharmaceutical compositions of peptide derivatized using an oxime linker
UA108994C2 (en) * 2009-11-17 2015-07-10 Ипсен Фарма С.А.С. Rormulation for hgh and rhigf-1 combination
KR101337797B1 (en) 2010-07-14 2013-12-06 한미사이언스 주식회사 A liquid formulation of long acting human growth hormone conjugate
WO2015046974A1 (en) * 2013-09-27 2015-04-02 한미약품 주식회사 Sustained type human growth hormone preparation
CN105764491A (en) 2013-12-09 2016-07-13 度瑞公司 Pharmaceutically active agent complexes, polymer complexes, and compositions and methods involving the same
US20160015789A1 (en) * 2014-07-17 2016-01-21 Teva Pharmaceutical Industries, Ltd. FORMULATIONS OF AN ALBUMIN hGH FUSION PROTEIN

Family Cites Families (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4783441A (en) * 1979-04-30 1988-11-08 Hoechst Aktiengesellschaft Aqueous protein solutions stable to denaturation
US4774091A (en) * 1983-10-14 1988-09-27 Sumitomo Pharmaceuticals Company, Ltd. Long-term sustained-release preparation
EP0189673B1 (en) * 1984-12-24 1990-09-26 Sumitomo Pharmaceuticals Company, Limited Stable growth hormone releasing factor preparation
PT83095B (en) * 1985-07-30 1987-12-23 Int Minerals & Chem Corp Method for the stabilization of growth promoting hormones using polyoxyethylene-polyoxypropylene copolymers
US4816568A (en) * 1986-05-16 1989-03-28 International Minerals & Chemical Corp. Stabilization of growth hormones
US4917685A (en) * 1986-05-16 1990-04-17 International Minerals & Chem. Corp. Delivery device for the administration of stabilized growth promoting hormones
ES2044941T5 (en) * 1987-08-21 1999-02-16 Mallinckrodt Group Inc STABILIZER OF GROWTH PROMOTING HORMONES.
US4837202A (en) † 1987-09-14 1989-06-06 Pitman-Moore, Inc. Method for stimulating the immune system
US5096885A (en) * 1988-04-15 1992-03-17 Genentech, Inc. Human growth hormone formulation
JPH01308235A (en) * 1988-06-03 1989-12-12 Yamanouchi Pharmaceut Co Ltd Human growth hormone for transnasal administration
US5008244A (en) * 1989-05-15 1991-04-16 Pitman-Moore, Inc. Method for increasing fertility in animals
CA2020654A1 (en) * 1989-07-07 1991-01-08 Yohko Akiyama Stabilized fgf composition and production thereof
IN172427B (en) * 1989-11-14 1993-07-24 Fidia Spa
US5217954A (en) * 1990-04-04 1993-06-08 Scios Nova Inc. Formulations for stabilizing fibroblast growth factor
US5126324A (en) * 1990-06-07 1992-06-30 Genentech, Inc. Method of enhancing growth in patients using combination therapy
WO1992017200A2 (en) * 1991-03-28 1992-10-15 Genentech, Inc. Stable growth hormone metal ion formulations
DK204791D0 (en) * 1991-12-20 1991-12-20 Novo Nordisk As UNKNOWN PHARMACEUTICAL PREPARATION
CA2636421C (en) † 1992-04-30 2010-06-29 Millennium Pharmaceuticals, Inc. A method of platelet and whole blood storage
ATE359824T1 (en) * 1992-07-31 2007-05-15 Genentech Inc AQUEOUS FORMULATION CONTAINING HUMAN GROWTH HORMONE

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6150331A (en) * 1998-02-04 2000-11-21 Jcr Pharmaceuticals Co., Ltd. Human growth hormone-containing aqueous pharmaceutical composition
JP2012051891A (en) * 1999-07-12 2012-03-15 Sandoz Ag Growth hormone formulation
JP2014208656A (en) * 1999-07-12 2014-11-06 サンド・アクチエンゲゼルシヤフト Growth hormone formulation
JP2015514704A (en) * 2012-03-30 2015-05-21 ハンミ サイエンス カンパニー リミテッドＨａｎｍｉ Ｓｃｉｅｎｃｅｃｏ．，Ｌｔｄ． High concentration solution of long-acting human growth hormone conjugate
JP2018115166A (en) * 2012-03-30 2018-07-26 ハンミ サイエンス カンパニー リミテッドＨａｎｍｉ Ｓｃｉｅｎｃｅ Ｃｏ．，Ｌｔｄ． Liquid formulation of highly concentrated long-acting human growth hormone conjugate
US10064951B2 (en) 2012-03-30 2018-09-04 Hanmi Science Co., Ltd. Liquid formulation of highly concentrated long-acting human growth hormone conjugate

Also Published As

Publication number Publication date
SE9201073D0 (en) 1992-04-03
NO310804B1 (en) 2001-09-03
AU3913593A (en) 1993-11-08
CA2102693A1 (en) 1993-10-04
DE69329367T2 (en) 2001-06-21
JP3268557B2 (en) 2002-03-25
US5567677A (en) 1996-10-22
EP0587858B2 (en) 2007-11-14
GR3034925T3 (en) 2001-02-28
NZ251498A (en) 1995-07-26
EP0587858B1 (en) 2000-09-06
CA2102693C (en) 2008-06-17
EP0587858A1 (en) 1994-03-23
PT587858E (en) 2001-02-28
NO934355L (en) 1993-12-01
AU666007B2 (en) 1996-01-25
DE69329367D1 (en) 2000-10-12
ES2150940T5 (en) 2008-04-16
DK0587858T3 (en) 2000-12-18
NO934355D0 (en) 1993-11-30
ATE196092T1 (en) 2000-09-15
ES2150940T3 (en) 2000-12-16
DE69329367T3 (en) 2008-06-19
WO1993019776A1 (en) 1993-10-14

Similar Documents

Publication Publication Date Title
JPH06508156A (en) Protein preparation consisting of growth hormone
AU733592B2 (en) A pharmaceutical formulation containing growth hormone, an amino acid and a non-ionic detergent
JPH03503764A (en) Human growth hormone preparation
JPH10511965A (en) Stabilized pharmaceutical preparation containing growth hormone and Leu-His-Leu
US8841252B2 (en) Pharmaceutical formulation
EP0785796B1 (en) A pharmaceutical formulation comprising a growth hormone and isoleucine
CA2311754C (en) A one dose syringe, comprising a freeze-dried protein composition, for administering of a volume less than 0.5 ml
JPH10507181A (en) Pharmaceutical formulation comprising growth hormone and valine
JPH10511964A (en) Stabilized pharmaceutical preparation containing growth hormone and X-Lys
CA2226553A1 (en) A stabilized pharmaceutical formulation comprising a growth hormone and a peptide comprising at least, one basic amino acid residue and at least one acid amino acid residue
WO1996021459A1 (en) A stabilized pharmaceutical formulation comprising a growth hormone and lys-x

Legal Events

Date Code Title Description
R250 Receipt of annual fees

Free format text: JAPANESE INTERMEDIATE CODE: R250

R250 Receipt of annual fees

Free format text: JAPANESE INTERMEDIATE CODE: R250

R250 Receipt of annual fees

Free format text: JAPANESE INTERMEDIATE CODE: R250

FPAY Renewal fee payment (event date is renewal date of database)

Free format text: PAYMENT UNTIL: 20080118

Year of fee payment: 6

FPAY Renewal fee payment (event date is renewal date of database)

Free format text: PAYMENT UNTIL: 20090118

Year of fee payment: 7

FPAY Renewal fee payment (event date is renewal date of database)

Free format text: PAYMENT UNTIL: 20090118

Year of fee payment: 7

FPAY Renewal fee payment (event date is renewal date of database)

Free format text: PAYMENT UNTIL: 20100118

Year of fee payment: 8

FPAY Renewal fee payment (event date is renewal date of database)

Free format text: PAYMENT UNTIL: 20110118

Year of fee payment: 9

FPAY Renewal fee payment (event date is renewal date of database)

Free format text: PAYMENT UNTIL: 20110118

Year of fee payment: 9

FPAY Renewal fee payment (event date is renewal date of database)

Free format text: PAYMENT UNTIL: 20120118

Year of fee payment: 10

FPAY Renewal fee payment (event date is renewal date of database)

Free format text: PAYMENT UNTIL: 20130118

Year of fee payment: 11